肿瘤坏死因子受体相关因子2对胃癌细胞生物学行为的影响及其机制  

The role and mechanism of TRAF2 in the biological behavior of gastric cancer

作　　者：戴菡珏 徐晶晶[2] 李文静[3] 国风 Dai Hanjue;Xu Jingjing;Li Wenjing;Guo Feng(Oncology Center,Changzhou Second People′s Hospital Affiliated to Nanjing Medical University,Changzhou 213003,China;Center for Clinical Laboratory,the First Affiliated Hospital of Soochow University,Suzhou 215006,China;Center for Clinical Laboratory,Nanjing Medical University Affiliated Suzhou Hospital,Suzhou 215001,China;Department of Oncology,Nanjing Medical University Affiliated Suzhou Hospital,Suzhou 215001,China)

机构地区：[1]南京医科大学附属常州第二人民医院肿瘤中心,213000 [2]苏州大学附属第一医院检验科,215006 [3]苏州市立医院检验科,215005 [4]苏州市立医院肿瘤科,215005

出　　处：《中华肿瘤杂志》2019年第11期805-812,共8页Chinese Journal of Oncology

基　　金：苏州市科技计划计划项目(SS201875);江苏省青年医学重点人才培养项目(QNRC2016725)。

摘　　要：目的探讨肿瘤坏死因子受体相关因子2(TRAF2)对胃癌AGS细胞生长、迁移和侵袭等细胞生物学行为的影响及机制。方法构建低表达TRAF2的胃癌AGS细胞株(AGS-siTRAF2),采用xCELLigence系统和细胞活力实验检测细胞的生长和增殖情况。采用流式细胞术检测细胞凋亡和细胞周期,采用xCELLigence系统和Transwell实验检测细胞的迁移和侵袭能力。采用Western blot法和TransAM法检测低表达TRAF2对AGS细胞核转录因子κB(NF-κB)信号通路的影响。采用免疫组化法检测胃癌组织中TRAF2的表达,分析TRAF2表达与胃癌临床病理特征的关系和对患者生存的影响。结果xCELLigence系统动态检测结果显示,从8 h开始,AGS-siTRAF2细胞的生长速度明显慢于AGS-sictrl细胞。细胞活力实验显示,培养24、48和72 h时,AGS-siTRAF2细胞的吸光度(A)值分别为51296.00±2631.06、68389.25±6703.21和65559.50±6339.22,均显著低于AGS-sictrl细胞(均P<0.001)。流式细胞术检测结果显示,培养24、48和72 h时,AGS-siTRAF2细胞凋亡率分别为(1.42±0.07)%、(2.98±0.11)%和(1.56±0.03)%,均显著高于AGS-sictrl细胞(均P<0.05);AGS-siTRAF2细胞S期占(23.57±1.12)%,对照组为(19.49±1.19)%,差异有统计学意义(P=0.012)。xCELLigence系统检测显示,从3 h开始,AGS-siTRAF2细胞的迁移能力低于AGS-sictrl细胞。Transwell实验检测结果显示,培养24 h后,AGS-sictrl的细胞数为(121.7±6.7)个,AGS-siTRAF2的细胞数为(84.0±6.6)个,AGS-siTRAF2细胞的迁移细胞数明显少于AGS-sictrl细胞(P=0.002)。xCELLigence系统检测结果显示,从3 h开始,AGS-siTRAF2细胞的细胞指数低于AGS-sictrl细胞。Transwell实验检测结果显示,培养24 h,AGS-sictrl的细胞数为(109.3±3.1)个,AGS-siTRAF2的细胞数为(79.0±6.2)个,AGS-siTRAF2细胞的穿膜细胞数明显少于AGS-sictrl细胞(P=0.002)。Western blot检测结果显示,AGS-siTRAF2细胞中RelA、RelB、p50和p52表达水平均明显低于AGS-sictrl细胞。TransAM检测结果显示,AGS-siObjective To clarify the effect of TRAF2 in the biological behavior of gastric cancer and explore the mechanism.Methods TRAF2 stably depleted AGS cell was established.Cell growth was monitored by x-CELLigence system.Cell proliferation was detected using cell viability assay.The apoptosis and cell cycle were detected by flow cytometry.The difference of migration and invasion abilities were measured by real-time xCELLigence system and Transwell.The expression and activity of NF-κB signaling pathway were measured by western blot and TransAM assay.The expression of TRAF2 in gastric cancer tissue and its clinical significance were detected by immunohistochemistry.Results The cell index of AGS-siTRAF2 cells was significantly lower than that of AGS-sictrl cells at 8 h.In the cell viability assay,the A values of AGS-siTRAF2 cells were 51296.00±2631.06,68389.25±6703.21 and 65559.50±6339.22 at 24 h,48 h and 72 h.The values of the viability of AGS-siTRAF2 cells were significantly lower than those of AGS-sictrl cells(P<0.001).The results of flow cytometry showed that the apoptosis rates of AGS-siTRAF2 cells were(1.42±0.07)%,(2.98±0.11)%and(1.56±0.03)%at 24 h,48 h and 72 h,respectively,which were significantly higher than those of AGS-sictrl cells(all P<0.05).The distribution of S phase in AGS-siTRAF2 cells was(23.57±1.12)%,while that in the AGS-sictrl cells was(19.49±1.19)%.The difference was statistically significant(P=0.012).AGS-siTRAF2 cells migrated much slower than AGS-sictrl cells from 3 h and the number of migrated AGS-sictrl cells was 121.7±6.7 while that of AGS-siTRAF2 cells was 84.0±6.6(P=0.002).The cell index of AGS-siTRAF2 cells was less than that of AGS-sictrl cells from 3 h.In Transwell assay,the number of invaded AGS-sictrl cells was 109.3±3.1 after 24 h of culture,significantly higher than 79.0±6.2 of AGS-siTRAF2 cells(P=0.002).Western blot analysis showed that the expression levels of RelA,RelB,p50 and p52 in AGS-siTRAF2 cells were significantly lower than those in AGS-sictrl cells.The activitie

关 键 词：胃肿瘤 肿瘤坏死因子受体相关因子2 细胞生长 迁移 侵袭 核转录因子KB 

分 类 号：R73[医药卫生—肿瘤]